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Related Experiment Videos

A high-throughput Arabidopsis reverse genetics system.

Allen Sessions1, Ellen Burke, Gernot Presting

  • 1Torrey Mesa Research Institute, Syngenta, 3115 Merryfield Row, San Diego, California 92121, USA. allen.sessions@syngenta.com

The Plant Cell
|December 7, 2002
PubMed
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This summary is machine-generated.

Researchers created the Syngenta Arabidopsis Insertion Library (SAIL) with over 50,000 T-DNA insertion lines. This resource aids functional genomics research by providing mapped insertion sites for Arabidopsis thaliana.

Area of Science:

  • Plant molecular biology
  • Functional genomics
  • Arabidopsis thaliana research

Background:

  • Functional genomics requires efficient tools to understand gene function.
  • T-DNA insertion lines are valuable for creating gene knockouts and studying gene function in plants.
  • Previous methods for characterizing T-DNA insertions were less efficient for large-scale studies.

Purpose of the Study:

  • To develop a high-throughput method for generating and characterizing T-DNA insertion lines in Arabidopsis.
  • To create a comprehensive resource of mapped T-DNA insertion sites for the scientific community.
  • To facilitate functional analysis of Arabidopsis genes through identified insertion mutations.

Main Methods:

  • Generation of approximately 100,000 transformed Arabidopsis lines.

Related Experiment Videos

  • Development of a modified, high-throughput thermal asymmetric interlaced (TAIL)-PCR protocol.
  • Sequencing of 85,108 TAIL-PCR products and mapping T-DNA insertion sites against the Arabidopsis genome.

    • Main Results:

    • Successfully mapped T-DNA insertion sites in 52,964 individual Arabidopsis lines.
    • Identified a bias in T-DNA insertions, with fewer insertions found in predicted coding sequences.
    • Confirmed predicted T-DNA insertions in 76% (257/340) of tested lines using PCR assays.

    Conclusions:

    • The Syngenta Arabidopsis Insertion Library (SAIL) provides a valuable, large-scale resource for Arabidopsis functional genomics.
    • The developed TAIL-PCR protocol enables efficient characterization of T-DNA insertion sites.
    • The SAIL resource is publicly available to accelerate plant research and gene function discovery.